Difference between revisions of "Part:BBa K3866021"
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<p>The production of butyrate by bacteria can be possible with a set of genes. This part includes the promoter ParaBAD with RBS(baba), the superpart 1(baba) and the double terminator(baba). The superpart 1 has only four of the nine genes that catalyze the formation of butyric acid. Those are PhaA, PhaB, Crt and Ter genes. | <p>The production of butyrate by bacteria can be possible with a set of genes. This part includes the promoter ParaBAD with RBS(baba), the superpart 1(baba) and the double terminator(baba). The superpart 1 has only four of the nine genes that catalyze the formation of butyric acid. Those are PhaA, PhaB, Crt and Ter genes. | ||
| − | [[File:T--Thessaly-- | + | [[File:T--Thessaly--ParaBAD.Operon1.tt.png|700px|thumb|none|<i><b>Fig.2:</b>The alpha2 vector that includes the transcriptional unit of the Operon 1 with the ParaBAD promoter and a double terminator. The operon 1 consists of the PhaA, PhaB, Crt and Ter genes, responsible for the butyrate production.</i>]] |
===Usage and Biology=== | ===Usage and Biology=== | ||
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===References=== | ===References=== | ||
| − | + | *Miscevic D, Srirangan K, Kefale T, Abedi D, Moo-Young M, Chou CP. Production of cellulosic butyrate and 3-hydroxybutyrate in engineered Escherichia coli. Appl Microbiol Biotechnol. 2019 Jul;103(13):5215-5230. doi: 10.1007/s00253-019-09815-x. Epub 2019 May 2. PMID: 31049621. | |
| + | *Kallunki, Barisic, Jäättelä and Liu, 2019. How to Choose the Right Inducible Gene ExpressionSystem for Mammalian Studies?. Cells, 8(8), p.796. | ||
Latest revision as of 10:49, 29 September 2021
ParaBAD:RBS-Superpart 1-double terminator
The production of butyrate by bacteria can be possible with a set of genes. This part includes the promoter ParaBAD with RBS(baba), the superpart 1(baba) and the double terminator(baba). The superpart 1 has only four of the nine genes that catalyze the formation of butyric acid. Those are PhaA, PhaB, Crt and Ter genes.
Usage and Biology
First, PhaA and BktB are two β-ketothiolases that carry out the homo-fusion of two acetyl-CoA moieties into acetoacetyl-CoA. Then, the acetoacetyl-CoA is reduced to (S)-3-hydroxybutyryl-CoA (3-HB-CoA) by NADH-dependent Hbd or (R)-3-HB-CoA by NADPH-dependent PhaB. Both (R)-3-HB-CoA and (S)- 3-HB-CoA can be converted to (R)-3-HB and (S)-3-HB, respectively, by a CoA-removing enzyme, such as Pct, TesB, or Ptb/Buk. Alternatively, (R)-3-HB-CoA and (S)-3-HB-CoA can be converted to crotonyl-CoA by PhaJ and Crt, respectively. Crotonyl-CoA is then reduced to butyryl-CoA by NADH-dependent Ter, and subsequently converted to butyrate by one of the above CoA-removing enzymes.
Design Notes
The coding sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in alpha 2 vector BBa_K3505009 and has overhangs compatible for GoldenBraid cloning.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1608
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1608
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 4566
Illegal BamHI site found at 1148 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1608
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1608
Illegal NgoMIV site found at 1373
Illegal NgoMIV site found at 1477
Illegal AgeI site found at 983 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 965
Source
Synthesized by Twist Biosciences.
References
- Miscevic D, Srirangan K, Kefale T, Abedi D, Moo-Young M, Chou CP. Production of cellulosic butyrate and 3-hydroxybutyrate in engineered Escherichia coli. Appl Microbiol Biotechnol. 2019 Jul;103(13):5215-5230. doi: 10.1007/s00253-019-09815-x. Epub 2019 May 2. PMID: 31049621.
- Kallunki, Barisic, Jäättelä and Liu, 2019. How to Choose the Right Inducible Gene ExpressionSystem for Mammalian Studies?. Cells, 8(8), p.796.